title: Receptor for advanced glycation end products is subjected to protein ectodomain shedding by metalloproteinases.
abstract: The receptor for advanced glycation end products (RAGE) is a 55-kDa type I membrane glycoprotein of the immunoglobulin superfamily. Ligand-induced up-regulation of RAGE is involved in various pathophysiological processes, including late diabetic complications and Alzheimer disease. Application of recombinant soluble RAGE has been shown to block RAGE-mediated pathophysiological conditions. After expression of full-length RAGE in HEK cells we identified a 48-kDa soluble RAGE form (sRAGE) in the culture medium. This variant of RAGE is smaller than a 51-kDa soluble version derived from alternative splicing. The release of sRAGE can be induced by the phorbol ester PMA and the calcium ionophore calcimycin via calcium-dependent protein kinase C subtypes. Hydroxamic acid-based metalloproteinase inhibitors block the release of sRAGE, and by RNA interference experiments we identified ADAM10 and MMP9 to be involved in RAGE shedding. In protein biotinylation experiments we show that membrane-anchored full-length RAGE is the precursor of sRAGE and that sRAGE is efficiently released from the cell surface. We identified cleavage of RAGE to occur close to the cell membrane. Ectodomain shedding of RAGE simultaneously generates sRAGE and a membrane-anchored C-terminal RAGE fragment (RAGE-CTF). The amount of RAGE-CTF increases when RAGE-expressing cells are treated with a gamma-secretase inhibitor, suggesting that RAGE-CTF is normally further processed by gamma-secretase. Identification of these novel mechanisms involved in regulating the availability of cell surface-located RAGE and its soluble ectodomain may influence further research in RAGE-mediated processes in cell biology and pathophysiology.
authors.affiliation: Institute of Biochemistry, Johannes Gutenberg University of Mainz, Becherweg 30, Mainz D-55099, Germany.
authors.forename: Ling
authors.initials: L
authors.lastname: Zhang
authors.forename: Monika
authors.initials: M
authors.lastname: Bukulin
authors.forename: Elzbieta
authors.initials: E
authors.lastname: Kojro
authors.forename: Annette
authors.initials: A
authors.lastname: Roth
authors.forename: Verena V
authors.initials: VV
authors.lastname: Metz
authors.forename: Falk
authors.initials: F
authors.lastname: Fahrenholz
authors.forename: Peter P
authors.initials: PP
authors.lastname: Nawroth
authors.forename: Angelika
authors.initials: A
authors.lastname: Bierhaus
authors.forename: Rolf
authors.initials: R
authors.lastname: Postina
mesh_terms: D051722:ADAM Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D000072197:ADAM10 Protein; D017398:Alternative Splicing / Q000187:drug effects / Q000235:genetics; D000544:Alzheimer Disease / Q000235:genetics / Q000378:metabolism; D053829:Amyloid Precursor Protein Secretases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D000001:Calcimycin / Q000494:pharmacology; D002273:Carcinogens / Q000494:pharmacology; D002460:Cell Line; D002462:Cell Membrane / Q000235:genetics / Q000378:metabolism; D048909:Diabetes Complications / Q000235:genetics / Q000378:metabolism; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D061965:Matrix Metalloproteinase Inhibitors; D008565:Membrane Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D011480:Protease Inhibitors / Q000494:pharmacology; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D017434:Protein Structure, Tertiary / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic / Q000235:genetics / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D002273:Carcinogens; D006877:Hydroxamic Acids; D007476:Ionophores; D061965:Matrix Metalloproteinase Inhibitors; D008565:Membrane Proteins; D011480:Protease Inhibitors; D020033:Protein Isoforms; D034741:RNA, Small Interfering; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic; D000001:Calcimycin; D053829:Amyloid Precursor Protein Secretases; D051722:ADAM Proteins; D020780:Matrix Metalloproteinase 9; D000072197:ADAM10 Protein; C491236:ADAM10 protein, human; D013755:Tetradecanoylphorbol Acetate